The invention relates to novel bicyclic lactam ring system containing compounds including their pharmaceutically acceptable isomers, salts, hydrates, solvates and prodrug derivatives, and pharmaceutically acceptable compositions thereof which are potent and highly selective inhibitors of isolated factor Xa or when assembled in the prothrombinase complex. These compounds show selectivity for factor Xa versus other proteases of the coagulation (e.g. thrombin, fVIIa, fIXa) or the fibrinolytic cascades (e.g. plasminogen activators, plasmin). In another aspect, the present invention relates to novel bicyclic lactam ring system containing compounds including their pharmaceutically acceptable isomers, salts, hydrates, solvates and prodrug derivatives, and pharmaceutically acceptable compositions thereof which are useful as potent and specific inhibitors of blood coagulation in mammals. In yet another aspect, the invention relates to methods for using these inhibitors as diagnostic or therapeutic agents for disease states in mammals characterized by undesired thrombosis or coagulation disorders.
Hemostasis, the control of bleeding, occurs by surgical means, or by the physiological properties of vasoconstriction and coagulation. The invention is particularly concerned with blood coagulation and ways in which it assists in maintaining the integrity of mammalian circulation after injury, inflammation, disease, congenital defect, dysfunction or other disruption. Under normal hemostatic circumstances, the body maintains an acute balance of clot formation and clot removal (fibrinolysis). The blood coagulation cascade involves the conversion of a variety of inactive enzymes (zymogens) into active enzymes which ultimately convert the soluble plasma protein fibrinogen into an insoluble matrix of highly cross-linked fibrin. Davie, E. J. et al., xe2x80x9cThe Coagulation Cascade: Initiation, Maintenance and Regulationxe2x80x9d, Biochemistry, 30, 10363-10370 (1991). These plasma glycoprotein zymogens include Factor XII, Factor XI, Factor IX, Factor X, Factor VII, and prothrombin. Blood coagulation follows either the intrinsic pathway, where all of the protein components are present in blood, or the extrinsic pathway, where the cell membrane protein tissue factor plays a critical role. Clot formation occurs when fibrinogen is cleaved by thrombin to form fibrin. Blood clots are composed of activated platelets and fibrin.
Blood platelets which adhere to damaged blood vessels are activated and incorporated into the clot and thus play a major role in the initial formation and stabilization of hemostatic xe2x80x9cplugsxe2x80x9d. In certain diseases of the cardiovascular system, deviations from normal hemostasis push the balance of clot formation and clot dissolution towards life-threatening thrombus formation when thrombi occlude blood flow in coronary vessels (myocardial infarctions) or limb and pulmonary veins (venous thrombosis). Although platelets and blood coagulation are both involved in thrombus formation, certain components of the coagulation cascade are primarily responsible for the amplification or acceleration of the processes involved in platelet aggregation and fibrin deposition.
Thrombin is a key enzyme in the coagulation cascade as well as in hemostasis. Thrombin plays a central role in thrombosis through its ability to catalyze the conversion of fibrinogen into fibrin and through its potent platelet activation activity. Under normal circumstances, thrombin can also play an anticoagulant role in hemostasis through its ability to convert protein C into activated protein C (aPC) in a thrombomodulin-dependent manner. However, in atherosclerotic arteries these thrombin activities can initiate the formation of a thrombus, which is a major factor in pathogenesis of vasoocclusive conditions such as myocardial infarction, unstable angina, nonhemorrhagic stroke and reocclusion of coronary arteries after angioplasty or thrombolytic therapy. Thrombin is also a potent inducer of smooth muscle cell proliferation and may therefore be involved in a variety of proliferative responses such as restenosis after angioplasty and graft induced atherosclerosis. In addition, thrombin is chemotactic for leukocytes and may therefore play a role in inflammation. (Hoover, R. J., et al. Cell, 14, 423 (1978); Etingin, O. R., et al., Cell, 61 657 (1990). These observations indicate that inhibition of thrombin formation or inhibition of thrombin itself may be effective in preventing or treating thrombosis, limiting restenosis and controlling inflammation.
Direct or indirect inhibition of thrombin activity has been the focus of a variety of recent anticoagulant strategies as reviewed by Claeson, G., xe2x80x9cSynthetic Peptides and Peptidomimetics as Substrates and Inhibitors of Thrombin and Other Proteases in the Blood Coagulation Systemxe2x80x9d, Blood Coag. Fibrinol. 5, 411-436 (1994). Several classes of anticoagulants currently used in the clinic directly or indirectly affect thrombin (i.e. heparins, low-molecular weight heparins, heparin-like compounds and coumarins).
The formation of thrombin is the result of the proteolytic cleavage of its precursor prothrombin at the Arg-Thr linkage at positions 271-272 and the Arg-Ile linkage at positions 320-321. This activation is catalyzed by the prothrombinase complex, which is assembled on the membrane surfaces of platelets, monocytes, and endothelial cells. The complex consists of Factor Xa (a serine protease), Factor Va (a cofactor), calcium ions and the acidic phospholipid surface. Factor Xa is the activated form of its precursor, Factor X, which is secreted by the liver as a 58 kd precursor and is converted to the active form, Factor Xa, in both the extrinsic and intrinsic blood coagulation pathways. Factor X is a member of the calcium ion binding, gamma carboxyglutamyl (Gla)-containing, vitamin K dependent, blood coagulation glycoprotein family, which also includes Factors VII and IX, prothrombin, protein C and protein S (Furie, B., et al., Cell, 53, 505 (1988)). The activity of Factor Xa in effecting the conversion of prothrombin to thrombin is dependent on its inclusion in the prothrombinase complex.
The prothrombinase complex converts the zymogen prothromnbin into the active procoagulant thrombin. It is therefore understood that Factor Xa catalyzes the next-to-last step in the blood coagulation cascade, namely the formation of the serine protease thrombin. In turn, thrombin then acts to cleave soluble fibrinogen in the plasma to form insoluble fibrin.
The location of the prothrombinase complex at the convergence of the intrinsic and extrinsic coagulation pathways, and the resulting significant amplification of thrombin generation (several hundred-thousand fold faster in effecting the conversion of prothrombin to thrombin than Factor Xa in soluble form) mediated by the complex at a limited number of targeted catalytic units present at vascular lesion sites, suggests that inhibition of thrombin generation is a desirable method to block uncontrolled procoagulant activity. It has been suggested that compounds which selectively inhibit factor Xa may be useful as in vitro diagnostic agents, or for therapeutic, administration in certain thrombotic disorders, see e.g., WO 94/13693. Unlike thrombin, which acts on a variety of protein substrates as well as at a specific receptor, factor Xa appears to have a single physiologic substrate, namely prothrombin.
Plasma contains an endogenous inhibitor of both the factor VIIa-tissue factor (TF) complex and factor Xa called tissue factor pathway inhibitor (TFPI). TFPI is a Kunitz-type protease inhibitor with three tandem Kunitz domains. TFPI inhibits the TF/fVIIa complex in a two-step mechanism which includes the initial interaction of the second Kunitz domain of TFPI with the active site of factor Xa, thereby inhibiting the proteolytic activity of factor Xa. The second step involves the inhibition of the TF/fVIIa complex by formation of a quaternary complex TF/fVIIa/TFPI/fXa as described by Girard, T. J. et al., xe2x80x9cFunctional Significance of the Kunitz-type Inhibitory Domains of Lipoprotein-associated Coagulation Inhibitorxe2x80x9d, Nature, 338, 518-520 (1989).
Polypeptides derived from hematophagous organisms have been reported which are highly potent and specific inhibitors of factor Xa. U.S. Pat. No. 4,588,587 describes anticoagulant activity in the saliva of the Mexican leech, Haementeria officinalis. A principal component of this saliva was shown to be the polypeptide factor Xa inhibitor, antistasin (ATS), by Nutt, E. et al., xe2x80x9cThe Amino Acid Sequence of Antistasin, a Potent Inhibitor of Factor Xa Reveals a Repeated Internal Structurexe2x80x9d, J. Biol. Chem., 263, 10162-10167 (1988).
Another potent and highly specific inhibitor of Factor Xa, called tick anticoagulant peptide (TAP), has been isolated from the whole body extract of the soft tick Ornithidoros moubata, as reported by Waxman, L., et al., xe2x80x9cTick Anticoagulant Peptide (TAP) is a Novel Inhibitor of Blood Coagulation Factor Xaxe2x80x9d Science, 248, 593-596 (1990).
Other polypeptide type inhibitors of factor Xa have been reported including the following: Condra, C. et al., xe2x80x9cIsolation and Structural Characterization of a Potent Inhibitor of Coagulation Factor Xa from the Leech Haementeria ghilianiixe2x80x9d, Thromb. Haemost., 61, 437-441 (1989); Blankenship, D. T. et al., xe2x80x9cAmino Acid Sequence of Ghilanten: Anti-coagulant-antimetastatic Principle of the South American Leech, Haementeria ghilianiixe2x80x9d, Biochem. Biophys. Res. Commun. 166, 1384-1389 (1990); Brankamp, R. G. et al., xe2x80x9cGhilantens: Anticoagulants, Antimetastatic Proteins from the South American Leech Haementeria ghilianiixe2x80x9d, J. Lab. Clin. Med., 115, 89-97 (1990); Jacobs, J. W. et al., xe2x80x9cIsolation and Characterization of a Coagulation Factor Xa Inhibitor from Black Fly Salivary Glandsxe2x80x9d, Thromb. Haemost., 64, 235-238 (1990); Rigbi, M. et al., xe2x80x9cBovine Factor Xa Inhibiting Factor and Pharmaceutical Compositions Containing the Samexe2x80x9d, European Patent Application, 352,903; Cox, A. C., xe2x80x9cCoagulation Factor X Inhibitor From the Hundred-pace Snake Deinagkistrodon acutus, venomxe2x80x9d, Toxicon, 31, 1445-1457 (1993); Cappello, M. et al., xe2x80x9cAncylostoma Factor Xa Inhibitor: Partial Purification and its Identification as a Major Hookworm-derived Anticoagulant In Vitroxe2x80x9d, J. Infect. Dis., 167, 1474-1477 (1993); Seymour, J. L. et. al., xe2x80x9cEcotin is a Potent Anticoagulant and Reversible Tight-binding Inhibitor of Factor Xaxe2x80x9d, Biochemistry 33, 3949-3958 (1994).
Factor Xa inhibitory compounds which are not large polypeptide-type inhibitors have also been reported including: Tidwell, R. R. et al., xe2x80x9cStrategies for Anticoagulation With Synthetic Protease Inhibitors. Xa Inhibitors Versus Thrombin Inhibitorsxe2x80x9d, Thromb. Res., 19, 339-349 (1980); Turner, A. D. et al., xe2x80x9cp-Amidino Esters as Irreversible Inhibitors of Factor IXa and Xa and Thrombinxe2x80x9d, Biochemistry, 25, 4929-4935 (1986); Hitomi, Y. et al., xe2x80x9cInhibitory Effect of New Synthetic Protease Inhibitor (FUT-175) on the Coagulation Systemxe2x80x9d, Haemostasis, 15, 164-168 (1985); Sturzebecher, J. et al., xe2x80x9cSynthetic Inhibitors of Bovine Factor Xa and Thrombin. Comparison of Their Anticoagulant Efficiencyxe2x80x9d, Thromb. Res., 54, 245-252 (1989); Kam, C. M. et al., xe2x80x9cMechanism Based Isocoumarin Inhibitors for Trypsin and Blood Coagulation Serine Proteases: New Anticoagulantsxe2x80x9d, Biochemistry, 27, 2547-2557 (1988); Hauptmann, J. et al., xe2x80x9cComparison of the Anticoagulant and Antithrombotic Effects of Synthetic Thrombin and Factor Xa Inhibitorsxe2x80x9d, Thromb. Haemost., 63, 220-223 (1990); Miyadera, A. et al., Japanese Patent Application JP 6327488; Nagahara, T. et al., xe2x80x9cDibasic (Amidinoaryl)propanoic Acid Derivatives as Novel Blood Coagulation Factor Xa Inhibitorsxe2x80x9d, J. Med. Chem., 37, 1200-1207 (1994); Vlasuk, G. P. et al., xe2x80x9cInhibitors of Thrombosisxe2x80x9d, WO 93/15756; and Brunck, T. K. et al., xe2x80x9cNovel Inhibitors of Factor Xaxe2x80x9d, WO 94/13693.
A number of inhibitors of trypsin-like enzymes (such as trypsin, enterokinase, thrombin, kallikrein, plasmin, urokinase, plasminogen activators and the like) have been the subject of disclosures. For example, Austen et al., U.S. Pat. No. 4,593,018 describes oligopeptide aldehydes which are specific inhibitors of enterokinase; Abe et al., U.S. Pat. No. 5,153,176 describes tripeptide aldehydes which have inhibitory activity against multiple serine proteases such as plasmin, thrombin, trypsin, kallikrein, factor Xa, urokinase, etc.; Brunck et al., European Publication WO 93/14779 describes substituted tripeptide aldehydes that are specific inhibitors of trypsin; U.S. Pat. Nos. 4,316,889, 4,399,065, 4,478,745 all disclose arginine aldehyde inhibitors of thrombin; Balasubramanian et al., U.S. Pat. No. 5,380,713 describes di- and tripeptide aldehydes which are useful for anti-trypsin and anti-thrombin activity; Webb et al., U.S. Pat. No. 5,371,072 describes tripeptide alpha-keto-amide derivatives as inhibitors of thrombosis and thrombin; Gesellchen et al., European Patent Publications 0479489 A2 and 0643073 A, describe tripeptide thrombin inhibitors; Veber et al., European Publication WO 94/25051 describes 4-cyclohexfylamine derivatives which selectively inhibit thrombin over other trypsin-like enzymes; Tapparelli et al., J. Biol. Chem. 268, 4734-4741 (1993) describe selective peptide boronic acid derivatives as inhibitors of thrombin.
Alternatively, agents which inhibit the vitamin K-dependent carboxylase enzyme, such as coumarin, have been used to treat coagulation disorders.
There exists a need for effective therapeutic agents for the regulation of hemostasis, and for the prevention and treatment of thrombus formation and other pathological processes in the vasculature induced by thrombin such as restenosis and inflammation.
The present invention provides novel bicyclic lactam ring system containing compounds including their pharmaceutically acceptable isomers, salts, hydrates, solvates and prodrug derivatives, which have particular biological properties and are useful as potent and specific inhibitors of blood coagulation in mammals. The invention also provides compositions containing such compounds. The compounds of the invention may be used as diagnostic reagents or as therapeutic agents for disease states in mammals which have coagulation disorders. Thus, the invention further provides methods for preventing or treating a condition in a mammal characterized by undesired thrombosis by administration of a therapeutically effective amount of a compound of the invention and a pharmaceutically acceptable carrier. Optionally, the methods of the invention comprise administering a pharmaceutical composition of the invention in combination with an additional therapeutic agent such as an antithrombotic and/or a thrombolytic agent and/or an anticoagulant. According to the invention, such conditions include, for example, any thrombotically mediated acute coronary or cerebrovascular syndrome, any thrombotic syndrome occurring in the venous system, any coagulopathy, and any thrombotic complications associated with extracorporeal circulation or instrumentation. The invention still further provides a method for inhibiting the coagulation of biological samples (e.g. stored blood products and samples).
The invention provides a compound of general formula I: 
wherein:
A is a member selected from the group consisting of: R2, xe2x80x94NR3R4, xe2x80x94C(xe2x95x90O)NR3R4, 
xe2x80x83where R2, R3, R4, R5, R6, R7, R8, and R9 are independently selected from the group consisting of H, xe2x80x94OH, C1-8alkyl, C2-8alkenyl, C2-8alkynyl, C3-8cycloalkyl, C6-12carbocyclic aryl, a five to ten membered heterocyclic ring system containing 1-4 heteroatoms selected from the group consisting of N, O and S; and C1-6alkylheterocyclic ring system having in the ring system 5 to 10 atoms with 1 to 4 of such atoms being selected from the group consisting of N, O and S; where R6 taken with either of R7 and R8, and/or R7 taken with R8, can each form a 5 to 6 membered heterocyclic ring containing from 1 to 4 atoms selected from the group consisting of N, O and S;
m is an integer from 0-3, preferably 0-2;
Z is a member selected from the group consisting of a direct link, C1-8alkyl, C3-8cycloalkyl, C2-8alkenyl, C2-8alkynyl, C1-8carbocyclic aryl, or a five to ten membered heterocyclic ring system containing 1-4 heteroatoms selected from the group consisting of N, O and S;
n is an integer from 0-3, preferably 0-2;
D is a member selected from the group consisting of a direct link, xe2x80x94CH2xe2x80x94, xe2x80x94Oxe2x80x94, xe2x80x94N(R2)xe2x80x94, xe2x80x94C(xe2x95x90O)xe2x80x94, xe2x80x94Sxe2x80x94, xe2x80x94SO2xe2x80x94, xe2x80x94SO2xe2x80x94N(R2)xe2x80x94, xe2x80x94N(R2)xe2x80x94SO2xe2x80x94, xe2x80x94OC(xe2x95x90O)xe2x80x94, xe2x80x94C(xe2x95x90O)Oxe2x80x94, xe2x80x94C(xe2x95x90O)xe2x80x94N(2)xe2x80x94 and xe2x80x94N(R2)xe2x80x94C(xe2x95x90O)xe2x80x94, where R2 is as described above;
R1 is a member selected from the group consisting of H, C1-8alkyl, C2-8alkenyl, C2-8alkynyl, C3-8cycloalkyl, halogen, polyhaloalkyl, C0-8alkyl-C(xe2x95x90O)OH, C0-8alkyl-C(xe2x95x90O)Oxe2x80x94C1-8alkyl, xe2x80x94CN, xe2x80x94NO2, C0-8alkyl-OH, C0-8alkyl-SH, xe2x80x94C(xe2x95x90O)NR2R3, xe2x80x94Oxe2x80x94R2 and xe2x80x94Oxe2x80x94C(xe2x95x90O)R2, an unsubstituted amino group, a mono or disubstituted amino group, wherein the substituted amino groups are independently substituted by at least one member selected from the group consisting of H, C1-8alkyl, C2-8alkenyl, C2-8alkynyl, C3-8cycloalkyl, polyhaloalkyl, xe2x80x94SO2R2, C0-8alkyl-C(xe2x95x90O)OH and C0-8alkyl-C(xe2x95x90O)Oxe2x80x94C1-8alkyl, where R2 and R3 is as described above;
q is an integer from 0-3, preferably 0-2;
X is N or xe2x80x94CR12;
R11 and R12 are independently a member selected from the group consisting of H, C1-8alkyl, C2-8alkenyl, C2-8alkynyl, C3-8cycloalkyl, C6-12carbocyclic aryl, C1-6alkylaryl, C1-6alkyl-C3-8cycloalkyl, xe2x80x94Oxe2x80x94R2, xe2x80x94Oxe2x80x94C(xe2x95x90O)R2, xe2x80x94C1-8alkyl-Oxe2x80x94R10, xe2x80x94C1-8alkyl-Oxe2x80x94C(xe2x95x90O)R10, xe2x80x94C1-8alkyl-C(xe2x95x90O)OR10, xe2x80x94C1-8alkyl-Oxe2x80x94C(xe2x95x90O)OR10, xe2x80x94C1-8alkyl-C(xe2x95x90O)NR10R10, xe2x80x94C1-8alkyl-NR10R10, xe2x80x94C1-8alkyl-NR10C(xe2x95x90O)R10, xe2x80x94SR10, where R2 is as described above and R10 is a member selected from the group consisting of H, C1-8alkyl, C2-8alkenyl, C2-8alkynyl, and wherein when two R10 groups are present they may be taken together to form a saturated or unsaturated ring with the atom to which they are both attached, preferably a partially or fully saturated ring;
p is an integer from 0-3, preferably 0-2;
E is a member selected from the group consisting of a direct link, xe2x80x94Oxe2x80x94, xe2x80x94N(xe2x80x94R11)xe2x80x94, where R11 is as set forth above, phenylene, a bivalent 5 to 12 member heteroaryl group containing 1 to 4 heteroatoms selected from the group consisting of N, O and S, and a five to ten membered nonaromatic bivalent heterocyclic ring system containing 1-4 heteroatoms selected from the group consisting of N, O and S, wherein said heteroaryl and said nonaromatic heterocyclic ring structure may be independently substituted by from 0 to 5 R14 groups and each R14 group is independently defined the same as the substituents set forth above for the R1 group;
J is a member selected from the group consisting of a direct link, a bivalent C3-8cycloalkyl group, phenylene, a 5 to 12 member bivalent heteroaryl group containing 1 to 4 heteroatoms selected from the group consisting of N, O and S, and a five to ten membered nonaromatic bivalent heterocyclic ring system containing 1-4 heteroatoms selected from the group consisting of N, O and S wherein said heteroaryl and said nonaromatic heterocyclic ring structure may be independently substituted by from 0 to 5 R14 groups and each R14 group is independently defined the same as the substituents set forth above for the R1 group;
G is a member selected from the group consisting of: H; xe2x80x94CN; xe2x80x94OR17; 
wherein
t is an integer from 0 to 6,
u is the integer 0 or 1, and R17, R18, R19, R20, R21, R22, R23, R24, R25 and R26 are independently selected from the group consisting of H, xe2x80x94OH, C1-8alkyl, C2-8alkenyl, C2-8alkynyl, C3-8cycloalkyl, C6-12carbocyclic aryl, a five to ten membered heterocyclic ring system containing 1-4 heteroatoms selected from the group consisting of N, O and S; and C1-6alkylheterocyclic ring system having in the ring system 5 to 10 atoms with 1 to 4 of such atoms being selected from the group consisting of N, O and S; where r18 taken with R19, R22 taken with either of R24 and R25, and R24 taken with R25, can each independently form a 5 to 6 membered heterocyclic ring containing from 1 to 4 atoms selected from the group consisting of N, O and S;
with the proviso that when G is H, xe2x80x94CN, xe2x80x94OR17, either E or J must contain at least one N atom;
and all pharmaceutically acceptable isomers, salts, hydrates, solvates and prodrug derivatives thereof.